DOI: 10.4103/jcot.jcot_5_26 ISSN: 3050-5763

Anthocyanin from Black Rice (Oryza sativa L./Kalabhat) Induces Apoptosis and Cell Cycle Arrest in Ehrlich Ascites Carcinoma Cells via Reactive Oxygen Species-Mediated Mitochondrial Pathway

Atish Barua, Mrinmoyee Mondal, Debapriya Roy, Priya Samanta, Shampa Pakhira, Souradeep Biswas, Jhinuk Basu, Ugir Hossain Sk, Subhadip Hajra, Prosenjit Saha
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Abstract

Background:

Anthocyanins are a class of flavonoids responsible for the red, purple, and blue pigmentation in black rice, fruits, and vegetables and are known for their potent antioxidant properties. In this study, we elucidate the efficacy and molecular mechanisms underlying the anticancer potential of black rice-derived anthocyanins.

Materials and Methods:

Anthocyanins were extracted from water-soaked Oryza sativa L. and quantified using high-performance liquid chromatography. Antioxidant activity was evaluated by the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay. Acute oral toxicity was assessed through hematological and biochemical analyses. Ehrlich ascites carcinoma (EAC) was induced intraperitoneally, and tumor-bearing mice were treated with anthocyanin extract for 10 days. Tumor burden, body weight, and survival were monitored. Mechanistic investigations were performed by measurement of intracellular reactive oxygen species (ROS) and lipid peroxidation, and assessment of mitochondrial membrane potential using the 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide assay, DNA fragmentation assay, caspase 3 activity, and acridine orange/ethidium bromide staining.

Results:

Black rice bran contained 170.03 mg anthocyanins per 100 g. The extract exhibited dose-dependent 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity (30%–75%) and produced no significant alterations in liver, kidney, or hematological parameters in normal mice. In EAC-bearing mice, anthocyanin treatment significantly ( P < 0.05) reduced viable tumor cell counts and improved survival, with the strongest effect observed at 15 mg/kg body weight doses. Furthermore, anthocyanin treatment induced a marked elevation in intracellular ROS levels and lipid peroxidation, mitochondrial membrane depolarization, caspase-3 activation, DNA ladder formation, and characteristic apoptotic morphology.

Conclusion:

Black rice-derived anthocyanins exhibit potent antitumor activity by inducing ROS-mediated mitochondrial apoptosis. These findings suggest their potential as promising natural chemoprotective agents against breast cancer.

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